The identification and characterization of particular alleles or of mutations have been historically important in every branch of biology and medicine. The study of mutations has contributed significantly to the understanding of the mechanisms and pathways of both normal physiological processes as well as disease pathogenesis. It has now moved on to the study of the relationships between protein structure and function, and correlation between genotype and disease phenotype.
Various developments in molecular genetics and biology have revolutionized the ability to analyze genes at a nucleotide sequence level. The emerging constraint on advances in molecular pathology appears to be the ability to correlate mutant genotype with disease phenotype.
Accordingly, there exists a need in the art to detect small mutations or polymorphisms, involving alterations to one or several bases in a nucleic acid sequence. More particularly, there exists a need in the art to selectively and preferentially produce copies of mutant nucleic acids in mixtures, said mixture possibly also containing the parental nucleic acid so that the mutant nucleic acids can be further analyzed. To these ends, methods should enable the rapid analysis of specific sequences, with decreasing requirements on sample quality and quantity, time, and manual effort.